Cyclopentanophenanthrenyl-oxazines



rate 3,063,990 Patented Nov.- 13, 1952 ice The present invention relatesto oxazine-type compounds, particularly to compounds of the formula inwhich the divalent radical of the formula in which formulae R representsa l7-polyhydro-cyclopentanophenanthrenyl radical, R represents hydrogenor functionally converted hydroxy, R stands for H(H), oxo orfunctionally converted x0 and n stands for one of the numbers 0, l, 2, 3or 4, or in which R stands for a 3-polyhydro-cyclopentanophenanthrenylradical, as well as for a 6-polyhydro-cyclopentanophenanthrenyl radicalor a l6-polyhydro-cyc1opentanophenanthrenyl radical, salts or quaternaryammonium compounds thereof, as well as process for the preparation ofsuch compounds.

The carbocyclic arylene radical of the previouslyshown formula may berepresented by a 1,2-phenylene radical, which may be unsubstituted ormay contain as substituents lower alkyl, e.g. methyl, ethyl and thelike, etherified hydroxy, particularly lower alkoxy, e.g. methoxy,ethoxy and the like, esterified hydroxy, particularly halogeno, e.g.fiuoro, chloro, bromo and the like, etherified mercapto, particularlylower alkyl-mercapto, e.g. methylmercapto, ethylmercapto and the like,nitro, amino, especially acylated amino, such as lower alkanoylamino,e.g. acetylamino, propionyl amino and the like, as well asN,N-disubstituted amino, such as N,N-di-lower alkyl-amino, e.g.N,N-dimethylamino, N,N-diethylamino and the like, halogeno-lower alkyl,e.g. trifluoromethyl, or any other substituents. In a 1,2-phenyleneradical of the formula 1 c \li/js M4 in which the carbon atomrepresenting position 1 is attached to the oxygen atom of the oxazinering, one or more than one of the same or of different substituents maybe attached to any of the available positions, but advantageously to the4-position of such 1,2-phenylene radical.

The carbocyclic arylene radical may also stand for a bicycliccarbocyclic arylene radical, particularly for a 1,2- naphthyleneradical, which may be unsubstituted or substituted by one or more thanone of the same or of different substituents; substituents are, forexample, those mentioned hereinbefore. In the 1,2-naphthalene radical ofthe formula in which the carbon atom representing the 2-position isattached to the oxygen atom of the oxazine ring, such substituents areadvantageously attached to the 4-p0sition, the 5-position, the6-position, the 7-position and/or the 8-position. A bicyclic carbocyclicarylene radical may also stand for a 2,3-naphthylene radical,particularly if the 1-position of such 2,3-naphthylene radical carries asubstituent.

Although the carbocyclic arylene radical in the above formula ispreferably a monocyclic or a bicyclic carbocyclic l,2-arylene radical,it may also stand forother polycyclic carbocyclic arylene radicals, suchas, for example, 1,2-phenanthrylene, 2,3-phenanthrylene,3,4-phenanthrylene and the like, whereby these radicals may beunsubstituted or contain as substituents one or more than one of thepreviously-mentioned substituents.

R representing a 17-polyhydro-cyclopentanophenanthrenyl radical, has thegeneral formula AAW '11 1 15 4 6 in which R, stands for hydrogen ormethyl, and R represents methyl, hydroxymethyl, functionally convertedhydroxy-methyl, such as esterified hydroxy-methyl or etherifiedhydroxy-methyl, oxo-methyl or functionally converted oxo-methyl. Thisabove-shown polyhydrocyclopentanophenanthrenyl radical is attachedthrough the 17-position directly or by way of one of thepreviously-shown carbon chains with the nitrogen atom of the oxazinenucleus.

The above-depicted 17-polyhydro-cyclopentanophenanthrenyl radical maycarry additional substituents, such as, for example, lower aliphatichydrocarbons, particularly lower alkyl, e.g. methyl, ethyl and the like;such radicals are primarily present in the 1-position, the 2-p'osition,the 6-position, the ll-position, the 12-position and/or the 16-position.Functional groups may also be attached to the17-polyhydro-cyclopentanophenanthrenyl radical; functional groups areparticularly oxygen-containing functional groups, such as hydroxy,functionally converted hydroxy, particularly esterified hydroxy and thelike, 0x0 or functionally converted oxo, particularly ketalized oxo andthe like.

An esterified hydroxy group, which may also represent the radical Rattached to one of the two-carbon atom chains previously mentioned, aswell as the esterified hydroxy portion in the functionally convertedhydroxymethyl group R mentioned hereinbefore, is primarily a hydroxygroup esterified with pharmaceutically acceptable organic carboxylicacids containing from one to twelve carbon atoms, such as aliphatichydrocarbon carboxylic acids, for example, alkanoic acids, primarilylower alkanoic acids, e.g. formic, acetic, propionic, butyric,

pivalic, heptanoic acid and the like, lower alkenoic acids, e.g.undecylenic acid and the like, cycloalkyl-lower alkane carboxylic acids,e.g. cyclohexylacetic, cyclopentylpropi- 'onic acid and the like,aliphatic hydrocarbon dicarboxylic acids, such as lower alkanedicarboxylic acids, e.g. succinic, glutaric acid, or halfesters of suchdicarboxylic acids with lower alkanol, e.g. methanol, ethanol and thelike, or cycloalkene dicarboxylic acids, e.g. tetrahydrophthalic acidand the like, carbocyclic aryl carboxylic acids, such as monocycliccarbocyclic aryl carboxylic acids, e.g. benzoic acid and the like,carbocyclic arylaliphatic hydrocarbon carboxylic acids, such asmonocyclic carbocyclic aryl-lower alkane carboxylic acids, e.g.phenylacetic, diphenylacetic, 3-phenylpropionic acid and the like, orany other suitable carboxylic acid. Hydroxy groups and hydroxy groupsesterified with carboxylic acids may advantageously be attached to the3-position, the 6-position, the 7-position, the ll-position, the12-position, the I l-position, the 16-position and/or the l7-position.

Other esterified hydroxy groups are, for example, those esterified withhydrohalic acids, which may be represented by halogeno atoms, e.g.fiuoro, chloro, bromo and the like; these halogeno atoms mayadvantageously be attached to the 2-position, the 4-position, the6-position, the 9-position, the l2-position and/or the 16-position. A

hydroxy group esterified by a mineral acid, Le. a halogeno atom, mayalso represent the group R attached to one of the two-carbon chainspreviously mentioned.

Oxo groups and/or ketalized X0 groups may be attached to the 3-position,the 7-position, the ll-position 'and/ or the l2-position; a ketalizedoxo group is primarily ketalized with a lower alkylene glycol, e.g.ethylene glycol, propylene glycol and the like. An oxo or functionallyconverted oxo group, as described hereinabove, may also represent thesubstituent R in one of the previouslyshown two-carbon side chains, aswell as the 0x0 or functionally converted oxo group in the oxo-methyl orfunctionally converted oxo-methyl group designated as R and attached tothe l3-position of the polyhydro-cyclopentanophenanthrenyl radical.

The polyhydro-cyclopentanophenanthrenyl radical may also contain doublebonds between neighboring carbon atoms; such double bonds maybe located,for example, at the l,2-'position, the 3,4-posit ion, the 4,5-position,the

j5,6-position, the 9,11-position, the 14,15-position and/ or the16,17-position.

The radical R may also stand for a 3-polyhydro-cyclopentanophenanthrenylnucleus of the formula 0x0. The 3-polyhydro-cyclopentanophenanthrenylnucleus may contain additional substituents, such as, for example, loweralkyl (particularly in the 1-position, the 2-position, the 6-position,the ll-position, the 12-position, the 16-p0sition and/or thel7-position), hydroxy or functionally converted hydroxy, particularlyesterified hydroxy (primarily attached to the 6-position, the7-position, the ll-position, the 12-position, the l4-position and/or thel6-position), including halogeno (substituting particularly the2-position, the 4-position, the 6-position, the 9-posi tion, thel2-position and/ or the l6-position), or an 0x0 or functionallyconverted oxo group (present, for example, in the 7-position, thell-position and/or the 12- position). Specific examples of suchsubstituents have been described hereinbefore. Double bonds may also bepresent, for example, in the 1,2-position, the 4,5-position, the9,11-position, the 14,15-position and/or the 16,17-position. Aspreviously mentioned, the 3-polyhydro-cyclopentanophenanthrenyl radicalis attached to the nitrogen atom of the oxazine portion through thecarbon atom representing the 3-position.

Other polyhydro cyclopentanophenanthrenyl radicals, which may representthe radical R, are the 7-polyhydrocyclopentanophenanthrenyl radical ofthe formula in which R R and R have the previously-given meaning, or anyother suitable polyhydro-cyclopentanophenanthrenyl nucleus, which maycontain additional sub- .stituents or double bonds as previouslymentioned.

The salts of the compounds of the present invention are primarilytherapeutically acceptable acid addition salts with inorganic acids,particularly mineral acids, eg. hydrochloric, hydrobromic, sulfuric,phosphoric acids and the like; with organic carboxylic acids, e.g.acetic, propionic, malonic, succinic, maleic, hydroxymaleic,dihydroxymaleic, melic, tartaric, citric, benzoic, mandelic acid and thelike, or with organic sulfonic acids, e.g. methane sultonic, ethanesulfonic, Z-hydroxyethane sulfonic, p-toluene sulfonic acid and thelike.

Quarternary ammonium derivatives of the compounds of this invention areprimarily those formed with reactive esters of lower alkanols withstrong acids, such as those with lower alkyl halides, e.g. methyl,ethyl, n-propyl, isopropyl or n-butyl chloride, bromide or iodide andthe like, di-lower alkyl sulfates, e.g. dimethyl sulfate, diethylsulfate and the like, lower alkyl lower alkane sulfonates, e.g. methylor ethyl methane or ethane sulfonate and the like, or lower alkylcarbocyclic aryl sulfonates, e.g. methyl p-toluene sulfonate and thelike.

Compounds of the present invention have antibacterial ellects againstgram-positive organisms, such as, for ex- 5 ample, Staph. aurezrs, B.szrbtilz's and the like, or gramnegative bacteria, such as, for example,E. coli, Ps. aeruginosa and the like, as well as antifungal activitiesagainst fungi, such as, for example, Nocardia asteroides, Trichophytonmentagrophytes, Histoplasma capsulatum, 5 Blastomyces dermatitidis andthe like. These compounds can, therefore, be used as antibacterialand/or anti-fungal agents to counteract infections caused, for example,by the abovementioned microoganisms.

Particularly outstanding antibacterial and antifungal 10 effects areexerted by compounds of the formula in which R is represented by one ofthe groups of the formulae OH(H), OAc(H), in which Ac represents theacyl radical of a lower alkanoic 301d, e.g., acetic, propionic, pivalicacid and the like, as well as by oxygen or lower alkylenedioxy, and inwhich the group of the formula may he represented by the group of theformula 1 5 W i/ 5 CH2 3 in which R stands for lower alkoxy, e.g.methoxy,

ethoxy and the like, halogeno, e.g. chloro, bromo and the like, nitro,N-lower alkanoyl-amino, e.g. N-acetylamino,

N-propionylamino and the like, and is primarily attached to the4-position, or by the group of the formula CHt in which R representshydrogen or has the same meaning as A,, the therapeutically acceptableacid addition 75 6 salts, particularly with mineral acids, and the loweralkyl quaternary ammonium derivatives of such compounds.

Another group of compounds having outstanding antibacterial andantifungal effects are those of the formula on OH; 0

in which R the radical of the formula and the group of the formula havethe previously-given meaning, the therapeutically acceptable acidaddition salts, particularly with mineral acids, andthe lower alkylquaternary ammonium derivatives of such compounds.

The invention may also be represented, for example, by the compounds ofthe formula in which the group of the formula has the previously-givenmeaning, and in which R represents the group of the formula-OH'(H) and Rstands for H(H) or OH(H), the radical of the formula represents one ofthe groups of the formula o nlr o og CH and in which 11 stands for oneof the whole numbers 0, 1 or 2, the therapeutically acceptable acidaddition salts, particularly with mineral acids, and the lower alkylquaternary ammonium derivatives of such compounds.

'7 An additional group of very active compounds are those of the formulain which the group of the formula and the group of the formula have thepreviously-given meaning, R stands for hydrogen or lower alkanoyl and Rrepresents hydrogen or lower alkyl, the therapeutically acceptable acidaddition salts, particularly with mineral acids, or the lower alkylquaternary ammonium derivatives of such compounds.

The new compounds of this invention may be used in the form ofpharmaceutical preparations, which contain the new steroid compounds,the salts or the quaternary ammonium compounds thereof in admixture witha pharmaceutical organic or inorganic, solid or liquid carrier suitablefor enteral or parenteral administration. For making up the preparationsthere can be employed substances which do not react with the newcompounds, such as water, gelatine, lactose, starches, stearic acid,magnesium stearate, stearyl alcohol, talc, vegetable oils, benzylalcohols, gums, propylene glycol, polyalkylene glycols or any otherknown carrier for medicaments. The pharmaceutical preparations may be insolid form, for example, as capsules, tablets, dragees and the like, orin liquid form, for example, as solutions, suspensions, emulsions andthe like. If desired, they may contain auxiliary substances, such aspreserving, stabilizing, wetting, emulsifying agents and the like, saltsfor varying the osmotic pressure or bufiers. They may also contain, incombination, other therapeutically useful substances.

The compounds of this invention can also be used as intermediates in thepreparation of other, particularly pharmaceutically, useful compounds.

The compounds of the present invention may be prepared, for example, byreacting a phenolic compounds of the formula g X I H. in which thedivalent radical of the formula has the previously-given meaning, withan amine of the formula R-NH in which R has the previously-givenmeaning, and with at least two equivalent amounts of formaldehyde or areactive derivative thereof, and/or, if desired, converting a resultingsalt into a free compound, and/or, if desired, converting a resultingcompound into a salt or a quaternary ammonium compound thereof.

The reaction is preferably carried out in the presence of diluents,particularly Water-miscible, polar solvents, for example, alcohols, suchas lower alkanols, e.g. methanol, ethanol, n-propanol, isopropanol andthe like, others, e.g. tetrahydrofuran, p-dioxane, diethyleneglycoldimethylether and the like, ketones, e.g. acetone and the like,carboxylic acid amides, e.g., formamide, N,N-dimethylformamide and thelike, or water or mixtures of such solvents, as well as less polarsolvents, such as aliphatic hydrocarbons, e.g. hexane and the like, canbocyclic aryl hydrocarbons, e.g. benzene, toluene and the like, or anyother suitable solvent-s. Normally, the amine and the phenol are givento the solution of the formaldehyde, which may be used as an aqueoussolution thereof, or in the form of a polymer thereof, e.g.paraformaldehyde, trioxymethylene and the like, or of an acetal thereofWith a lower alkanol, e.g. methanol, ethanol and the like, such as adi-lower alkoxy-methane, e.g. dimethoxymethane, diethoxymethane and thelike. The reaction may be carried out in the presence of a condensingreagent, for example, a base, such as an alkali metal hydroxide, e.g.lithium hydroxide, sodium hydroxide, potassium hydroxide and the like.If necessary, the reaction mixture may be heated, for example, on thesteam bath to about S090, if desired, in the atmosphere of an inert gas,e.g. nitrogen.

The resulting benzoxazine compound may crystallize upon cooling, or itmay be isolated by diluting the solution with a solvent differing fromthe one used in the reaction and, if necessary, extracting the product,or by evaporating the solution and crystallizing the residue. Thereaction product is purified according to known methods, for example, bycrystallization, recrystallization, adsorption, for example, on aluminumoxide or any other suitable adsorbent, and elution, or any othersuitable method.

The starting materials used in the above reaction are known, or, if new,may be prepared according to known procedures.

The compounds of this invention may be obtained in the form of the freebases or as the salts thereof. A salt may be converted into the freebase, for example, by reaction with an alkaline reagent, such as analkali metal hydroxide, e.g. lithium, sodium or potassium hydroxide, analkali metal carbonate, e.g. sodium or potassium carbonate or hydrogencarbonate, ammonia, an ion exchange resin or any other suitable reagent.A free base may be converted into its therapeutically useful acidaddition salts by reaction with one of the inorganic or organic acidsoutlined hereinbefore; for example, a solution of the free base in asolvent, such as a lower alkanol, e.g. methanol, ethanol, propanol,isopropanol and the like, a monocyclic carbocyclic aryl hydrocarbon,e.g. benzene, toluene and the like, an ether, e.g. diethylether and thelike, a halogenated aliphatic hydrocarbon, e.g. methylene chloride andthe like, or in a mixture of such solvents may be reacted with the acidor a solution thereof and the desired salt may then be isolated.

Quaternary ammonium derivatives of the compounds of this invention maybe obtained, for example, by reacting the tertiary base with an esterformed by a lower alkanol and a strong inorganic or organic acid. Suchacids are more especially mineral acids, e.g. hydrochloric, hydrobromic,hydriodic, sulfuric acid and the like, or strong organic sulfonic acids,for example, lower alkane sulfonic acids, e.g. methane sulfonic acid,ethane sulfonic acid and the like, hydroxy-lower alkane sulfonic acids,e.g. Z-hydroxy-ethane sulfonic acid and the like, or carbocyclic arylsulfonic acid, e.g. p-toluene sulfonic acid and the like. Usefulreactive esters are, for example, lower alkyl halides, e.g. methyl,ethyl, n-propyl or isopropyl chloride, bromide or iodide and the like,di-lower alkyl sulfates, e.g. dimethyl sulfate, diethyl sulfate and thelike, lower alkyl lower alkane sulfonates, e.g. methyl or ethyl methaneor ethane sulfonate and the like, lower alkyl hydroxy-lower alkanesulfonates, e.g. methyl 2- room temperature or at an elevatedtemperature, at

atmospheric pressure or in a closed vessel under pressure, and ifdesired, in the atmosphere of an inert gas, e.g. nitrogen. Suitablesolvents are more especially lower alkanols, e.g. methanol, ethanol,propanol, isopropanol, butanol, pentanol and the like, lower alkanones,e.g. acetone, methyl ethyl ketone and the like, organic acid amides,e.g. formamide, dimethylformamide and the like, monocyclic carbocyclicaryl hydrocarbons, e.g. benzene and the like, halogenated hydrocarbons,e.g. methylene chloride and the like, ethers, e.g. diethyl ether and thelike, or any other equivalent solvent.

The invention also comprises any modification of the process wherein acompound obtainable as an intermediate at any stage of the process isused as starting material and the remaining step(s) of the processis(are) carried out, as well as any new intermediates.

In the process of this invention such starting materials are preferablyused which lead to final products mentioned in the beginning aspreferred embodiments of the invention.

The following examples are intended to illustrate the invention and arenot to be construed as being limitations thereon. Temperatures are givenin degrees centigrade.

Example 1 A trace of potassium hydroxide and 0.9 g. of tr-ioxymethyleneare dissolved in 75 ml. of anhydrous n-propanol while warming. Thesolution is cooled in ice, 4.8 g. of17-amino-5-androstene-3fi-ol[Ruzicka et al., Helv. Chirn. Act., vol. 19,p. 107 (1936)] is added, followed by 2.27 g. of4-(N-acetylamino)-phenol. The solution is refluxed under an atmosphereof nitrogen for fifteen hours and is then concentrated to dryness. Theresidue is suspended in 50 ml. of benzene, the insoluble material isfiltered off and recrystallized from methanol. This product is digestedwith 250 ml. of chloroform, the insoluble material is filtered ofi andrecrystallized from ethanol to yield the6-(N-acetylamino)-2,4-dihydro-3-( 3 8-hydroxy-S-androsten-l7-yl)-1,3-benzoxazine of the formula Example 2 To acold solution of 0.38 g. of trioxymethylene and a trace of potassiumhydroxide in ml. of methanol is added 2.0 g. of 17-amino-androstane-33-ol in 40 ml. of methanol, followed by 9.10 g. of S-naphthol. Thereaction mixture is allowed to stand for one hour at room temperatureunder an atmosphere of nitrogen and is then refluxed for ninety minutes.A crystalline product pre 10 cipitates on chilling, which is filteredofi" and recrystallized from a mixture of methylene chloride and ethanolto yield the desired2,3-dihydro-2-(3fl-hydroxy-17-androstanyl)-lH-naphth[1,2-e]-m-oxazine ofthe formula CH1 H C 11:

3 Cn W 0 HO which melts at 227-228"; yield: 2.56 g.

The starting material may be prepared as follows: To a refluxingsolution of 4.7 g. of 3fi-hydroxy-17-oximinoan'drostane -[Ruzicka etal., Helv. Chirn. Acta, vol. 17, p. 1395 (1934)] in ml. of propanol isadded slowly and in portions a total of 10.0 g. of sodium. The cooledsolution is poured into 3000 ml. of water, the precipitate is filteredoif, washed with Water and recrystallized several times from methanoland then from a mixture of methylene chloride and hexane to yield thedesired 17- amino 3B hydroxy-androstane, M.P. 163-164"; yield: 2.8 g.

The 2,3-dihydro-2-( 3 ,B-hydroxy-S-androsten-17-yl 1H-naphth[1,2-e]-rn-oxazine (M.P. 223-224"; yield: 79 percent), and2,3-dihydro-2-(3a-hydroxy 17 androstanyl)- 1H-naphth'[1,2-e]-m-oxazine(M.P. 176; yield: 54 percent) are prepared according to theabove-outlined procedure by selecting the appropriate startingmaterials.

Example 3 To a cold solution of 0.38 g. of trioxymethylene and a traceof potassium hydroxide in 25 m1. of anhydrous propanol is added 2.0 g.of 20w-amino-3p-hydroxy-5w pregnane, followed by 0.78 g. of4-methoxy-phenol. The reaction mixture is refluxed under an atmosphereof nitrogen for fifteen hours and then concentrated to dryness underreduced pressure; the residue is triturated with methanol and the solidmaterial is filtered off. The desired 2, 4-dihydro-3-(3,8-hydroxy-5a-pregnan-20a-yl)-6'- methoxy-1,3-benzoxazine ofthe formula melts at 125127 after repeated recrystallization from amixture of methylene chloride and methanol; yield: 1.1 g.

The starting material in the above reaction may be prepared as follows:A solution of 7.0 g. of SB-acetoxy- 20u-amino-5-pregnene, preparedaccording to the procedures of Julian et al., J. Am. Chem. Soc., vol.70, p. 888 (1948), in 100 ml. of a 2% methanol solution of potassiumhydroxide is refluxed for two hours. The reaction mixture is poured intoan excess of water and the aqueous solution extracted with ether. Theether solution is washed with water, dried over sodium sulfate andthesolvent is removed, leaving 5.7 g. of 20a-amino-36-hydroxy-S-pregnenewhich, after recrystallization from ethyl acetate, melts at 172474"; [u]=-69.2 (in chloroform).

1 1 A solution of 3.65 g. of 20a-amino-SB-hydroxy-S-pregnene in 50 ml.of glacial acetic acid is treated with hydrogen under atmosphericpressure and in the presence of platinum oxide. Upon completion of thereduction the catalyst is filtered off, the filtrate is concentrated todryness under reduced pressure and the residue dissolved in warm aqueousmethanol, to which a small amount of 10 percent aqueous sodium hydroxidehas been added. The 20wamino-3,8-hydroxy-5-a-pregnane precipitates inWhite crystals and is recrystallized from a mixture of ethanol andwater, M.P. 171-173"; yield: 2.55 g.

By selecting the appropriate starting materials and following theabove-described procedure, the 2,4-dihydro- 3 (313hyd-roxy-Saregnan-ZOIS-yl)-6-methoxy-1,3-benzoxazine (M.P. l55156 afterrepeated recrystallization from a mixture of methylene chloride andheptane; yield: 31 per cent) and the2,4-dihydro-3-(3,8-hydroxy-5-pregmen-203M)-6-methoxy-1,3'benzoxazine(MP. 152", after recrystallizaton from a mixture of methylene chlorideand methanol and drying at 110; yield: 21 percent) can be prepared.

The starting materials used for the preparation of the above-mentionedcompounds may be prepared as follows: A solution of 13.4 g. of3,8-hyd-roxy-20-oximino-5a-pregnane (prepared by treatment of a solutionof 3,8-hydroxy- 20-oxo-5a-pregnane in pyridine with hydroxylarninehydrochloride) in 750 ml. of dry n-propanol is refluxed while 25 g. ofsodium is added in portions over a period of forty-five minutes.Refluxing is continued until all the sodium is dissolved. The solutionis concentrated under reduced pressure to a volume of 350 ml., thendiluted with 3000 ml. of cold Water. The white, crystalline material isfiltered oft, washed with water and recrystallized from methanol toyield 8.5 g. of 2013-amino-3fi-hydroxy-Sa-pregnane.

To a refluxing solution of 11.39 g. of the oxime of3B-acetoxy-20-oxo-5-pregnene in 800 m1. of dry n-propanol is added 18 g.of sodium in small pieces over a period of three hours. About 300 ml. ofthe solvent is removed under reduced pressure and approximately 1000 ml.of water is added. The aqueous solution is extracted with ethyl acetate,the organic layer is Washed with water, dried over sodium sulfate andthen filtered. An ether solution of hydrogen chloride is added and 5.3g. of 20/3- amino-3fl-hydroxy-5-pregnene hydrochloride is obtained.

The salt is dissolved in warm ethanol by adding a small volume of water,and 10 percent aqueous sodium hydroxide is added to pH 8 to 9. Ondilution with water the 205-amino-35-hydroxy-5-pregnene precipitates, isfiltered oif and dried. After recrystallization from methanol and fromethyl acetate itmelts at 173174; yield: 4.2 g.

Example 4 To a solution of 0.39 g. of trioxymethylene and a trace ofpotassium hydroxide in 30 ml. of anhydrous n-propanol is added 2.1 g. of20B-amino-3fl-hydroxy-Spreg- CH3 CH2 12 is purified by recrystallizationfrom ethanol, MP. 249- 250; yield: 0.68 g.

The6-(N-acetylamino)-2,4-dihydro-3-(3,6-hydroxy-5epregnan-20a-yl)-1,3-benzoxazine(M.P. 247248"; yield: 29 percent) and 6-(N-acetylamino- 2,4dihydro-3-(3fi- -hydroxy-Sa-pregnan-ZOB-yl)-1,3-benzoxazine (MP. 245--247'; yield: 24 percent) are prepared according to the above-givenprocedure by selecting the appropriate starting material.

Example 5 1.0 g. of ZOfi-amino-S S-hydIoXy-Sa-pregnane in 15 ml. ofmethanol, followed by 0.45 g. of B-naphthol, is added to a cold solutionof 0.19 g. of trioxymethylene and a trace of potassium hydroxide in 5ml. of methanol. The reaction mixture is refluxed under an atmosphere ofnitrogen for two hours and is then chilled to yield the desired2,3-dihydro-2-(35 hydroxy-Sa-pIegnaH-ZOfl-yl)-1H-naphth[1,2-e]-m-oxazine of the formula Example 6 A solution of 1.0 g.of 23-amino-3a,12u-dihydroxynorcholane hydrochloride in 3 ml. ofmethanol is added 3.7 ml. of a 3.8 percent methanol solution ofpotassium hydroxide. The resulting potassium chloride is filtered offand 0.15 g. of trioxymethylene in 1 ml. of methanol containing a traceof potassium hydroxide, followed by 0.36 g. of S-naphthol is added. Thereaction mixture is refluxed under a nitrogen atmosphere for 1.5 hours,then concentrated to dryness under reduced pressure, the residue isdissolved in ethyl acetate, a small amount of diethyl ether is added andthe solution is filtered. The filtrate is concentrated under reducedpressure, the residue is dissolved in benzene and the desired2,3-dihydro- 2-(3a,12u-dihydroxy 23 norcholanyl)-1H-naphth[1,2- e]-m-oxazine of the formula is precipitated as the hydrochloride by addingdropwise a solution of hydrogen chloride in benzene. The hydro- 13chloride is recrystallized from a mixture of ethanol and ether, M.P.155156.

The starting material be prepared as follows: A solution of 18.0 g. of30L,lZCL-dlQCClOXY-HOICI'IOHC acid [Kazuno et al., J. Biochem (Japan),vol. 29, p. 421 (1939)] in 20 ml. of thionyl chloride is allowed tostand at room temperature for three hours and is then evaporated todryness under reduced pressure. Dry benzene is added, the solvent isevaporated under reduced pressure to remove remaining thionyl chloride.The residue is triturated with petroleum ether, the precipitate isfiltered off and added immediately to 50 ml. of liquid ammonia whilerapidly stirring. The ammonia is allowed to evaporate, the residue iswashed with Water and re crystallized from ethyl acetate to yield the3a,12a-diacetoxy-norcholic acid amide, M.P. 201-202; yield: 10.0 g.

A solution of 9.0 g. of the above amide in 90 ml. of dry tetrahydrofuranis added to 5.4 g. of lithium aluminum hydride in 900 ml. oftetrahydrofuran over a period of one hour. The reaction mixture isrefluxed for three hours, 30 ml. of water is cautiously added, followedby a small amount of powdered cellulose. The solid materials arefiltered oil, washed with ether and the filtrate is concentrated underreduced pressure. The residue is dissolved in chloroform, dry hydrogenchloride is passed through the solution, the solvent is decanted and theresidue is recrystallized from a mixture of ethanol and diethyl ether toyield the hydrochloride of 23-amino-3ot, 12m-dihydroxy-norcholane, M.P.300-302"; yield: 7.2 g.

Example 7 1.00 g. of 24-amino-3a,7a,12a-trihydroxy-cholane hydrochloride[Wessely et al., Monatsh. Chem, vol. 82, p. 437 (1951)] is convertedinto the free base by treatment of a methanol solution with methanolicpotassium hydroxide; after filtering oil the resulting potassiumchloride, 0.14 g. of trioxymethylene in 1 ml. of methanol containing asmall amount of potassium hydroxide and 0.34 g. of fl-naphthol areadded. The reaction mixture is treated and worked up as shown in Example6 toyield the hydrochloride of2,3-(llhydI'O-2-(3a,7oc,l-2oz-tllhydrxy-24-cholanyl)-1H-uaphth[1,2-e]-m-oxazineof the formula $113 2 I CH-CHRCHQGHPN on, I

HO OH M.P. 17l174 after recrystallization from a mixture of methanol anddiethyl ether.

Example 8 The potassium chloride, precipitating from a mixture of 10.0g. of 24-amino-3a,7a,l2a trihydroxy-cholane hydrochloride and 13.0 ml.of methanolic potassium hydroxide, is filtered oil, the filtrate iscooled in ice and a solution of 1.4 g. of trioxymethylene in ml. ofmethanol containing a trace of potassium hydroxide, followed by 4.00 g.of 4-brorno-phenolis added. The solution is refluxed under a nitrogenatmosphere for fifteen hours, is then cooled and concentrated to drynessunder reduced pressure. A solution of the residue in ml. of a 1:1-mixture of methylene chloride and benzene is chromatographed on 200 g.of aluminum oxide (Woelm, neutral activity II); the column is developedwith (a) 500 ml. of benzene, (b) 250 ml. of a lzl-mixture of methylenechloride and benzene, (c) methylene chloride and (d) 14 methylenechloride containing one percent of methanol. The (c)- and (d)- fractionsare combined and the solvents evaporated. The residue does notcrystallize; a benzene solution of the latter is added to cold petroleumether and the desired 6-bromo-2,4-dihydro-3-(3a,7x,l2utrihydroxy-24-cholanyl)1,3-benzoxazine of the formula CH CH2 7 HOBr HO --OH precipitates, yield: 3.2 g.

The 2,4-dihydro-6-methoxy-3-(3a,7a,12a trihydroxy-24-cholanyl)-1,3-benzoxazine (noncrystalliue; yield: 34 percent) and2,4-dihydro-6-methyl-3-(3u,7a,12a-trihydroxy-24-cholanyl) 1,3benzoxazine (non-crystalline; yield: 26 percent) are prepared accordingto the abovementioned procedure.

Example '9 0.5 g. of2,4-dihydro-3-(3fl-hydroxy-5-a-pregnan-20txyl)-6-methoxy-1,3-benzoxazineis dissolved in an excess of methyl iodide; the reaction mixture isallowed to stand for several days at room temperature. The excess methyliodide is distilled off under reduced pressure and the residue isrecrystallized from methanol to yield the methiodide of2,4-dihydro-3-(3p-hydroxy-Swpregnan-20ueyl)- 6-n1ethoxy-L3-benzoxazine.

Example 10 Hydrogen chloride gas is passed through a cold solution of0.8 g. of dihydro-3-(3fl-hydroxy-Sa-pregnan-ZOayl)-6-methoxy1,3-benzoxazine in benzene; the resulting 2,'4-'dihydro'3-(-3}8--hydroxy 5a pregnan-20a-yD-6-methoxy-1,3-benzoxazine precipitates, isfiltered off and recrlystallized from a cold mixture of ethanol anddiethyl et er.

What is claimed is:

1. 6 (N acetylamino)-2,4-dihydro-3-(3/3-hydroxy-5-androsten-17-yl)-1,3-benzoxazine.

2. 2,4 dihydro 3-(3p-hydroxy-5-androsten-17-yl)-6-methoxy-1,3-benzoxazine.

3. 2,3 dihydro 2 (3a-hydroxy-l7-androstanyl)11H-naphth[1,2-e]-m-oxazine.

4. 2,3 dihydro-2-(3;3-hydroxy-5-androsten-17-yl)-1H- naphth[ 1,2-e]-m-oxazine.

5. 2,3 dihydro 2 (3,6-hydroxy-17-androstanyl)-1H-naphth[1,2-e]-rn-oxazine.

6, 2,4 dihydro 3-(3p-hydroxy Sa-pregnan-ZOwyl)-6-methoxy-l-,3-benzoxazine.

8. 2,4 dihydro-3-(3fi-hydroxy 5 pregnen 20,8 yl)-6-methoxy-1,3-benzoxazine.

9. 6 N acetylamino)-2,4-dihydro-3 (3 3-hydroxy-5-pregnen-ZOfi-yl)-1,3-benzoxazine.

10. 6 (N acetylamino)-2,4-dihydro-3-(3,3-hydroxy- 5a-pregnan-20a-yl)-1,3-benzoxazine.

11. 6 (N :acetylamino)-2,4-dihydro-3-(3fl-hydroxy-5a-pregnan-20fi-yl)-1,3-benzoxazine.

12. 2,3 dihydro 2-(3fl-hydroxy-5a-pregnan-2OB-yl)-1H-naphth[1,2-e]:m-oxazine.

v13. 2,3 dihydro 2 (3p-hydroxy-Sa-pregnan-ZOa-yl)- 1H-naphth[ 1,2-e]m-oxazine.

14. 2,3 dihydro 2-(3B-hydroxy-S-pregnan-ZD S-yl)-lH-naphth[1,2-e]-m-oxazine.

15. 2,3 dihydro 2-(3a,12a-dihydroxy-23-norcholanyl) '1H-naphth 1,2-e]-m-oxazine.

in which R represents the group of the formula OH(H), and R stands for amember selected from the group consisting of the groups of the formulaeH(H) and OH(H), the radical of the formula:

I RBI represents one of the formulae selected firom the group consistingof and "n represents one of the whole numbers 0, 1 and 2, and in whicheach of the groups R-;' and R represents a member'selected from thegroup consisting of hydrogen, lower alkoxy, halogeno, nitro and N loweralkanoylamino, the therapeutically acceptable acid addition saltsthereof and the lower alkyl quaternary ammonium derivatives thereof.

21. A member selected from the group consisting of a compound having oneof therformulae:

o Hflll/ R7 are and

Ha R0 in which R is a member selected from the group consisting ofOH(H), OAc(H), in which Ac stands for lower alkanoyl, and loweralkylenedioxy, and the radical of the formula:

stands for one of the formulae selected from the group consisting of andand in which each of the groups R and R is a member selected from thegroup consisting of hydrogen, lower alkoxy, halogeno, nitro and N-loweralkanoylamino, the pharmacologically acceptable acid addition saltsthereof and the lower alkyl quaternary ammonium derivatives thereof.

22. A member selected from the group consisting of a compound having oneof the formulae:

in which R is a member selected from the group con- Slstmg of OH(H),OAC(H), m wh1ch Ac stands f and each of the groups R, and R is a memberselected lower alkanoyl, and lower alkylencdioxy, and the radical 15from the group consisting of hydrogen, lower alkoxy of the formula:halogeno, nitro and N-lower alkanoylarnino, the pharmak ceuticallyacceptable acid addition salts thereof and the alower alkyl quaternaryammonium derivatives thereof. stands for one of the formulae selectedfrom the group 20 consisting of No references cited.

UNITED STATES PATENT OFFICE CERTIFICATE OF CORRECTION Patent No, 35363990 1962 November 13 Martin Eric Kuehne It is hereby certified thaterror appears in the above numbered patent requiring correction and thatthe said Letters Patent should read as corrected below.

Column 1 lines ll to 22:

formulae reading "-=(CH (CH same column 15 lines 23 to 31 left-handportion of the formula reading line 3 for "brom02-4" read =-bromo2 4 andlines 23 to 31 for the portion of the each occurrence read for the lowercolumn 1e lines 3 to 13 the lower left=-hand portion of the formulashould appear as shown below instead of as in the patent:

Signed and sealed this 26th day of May 19640 SEAL Attest:

EDWARD Jo BRENNER Commissioner of Patents ERNEST W, SWIDER AttestingOfficer

20. A MEMBER SELECTED FROM THE GROUP CONSISTING OF COMPOUNDS OF THEFORMULAE: